Test carrier for analysis of a sample liquid, method of using a test carrier and process for making a test carrier layer

• US 5,071,746 A
• Filed: 04/13/1989
• Issued: 12/10/1991
• Est. Priority Date: 04/28/1988
• Status: Expired due to Fees

• Current Assignee: Boehringer Mannheim GmbH (Roche Holding AG)
• Original Assignee: Boehringer Mannheim GmbH (Roche Holding AG)
• Inventors: Lerch, Rolf, Wilk, Hans-Erich, Munter, Karin
• Primary Examiner(s): Nucker, Christine
• Assistant Examiner(s): STUCKER, JEFFREY J
• Application Number:
  US07/337,351
• Time in Patent Office: 971 Days
• Field of Search: 422/55, 422/56, 422/57, 436/514, 436/524, 436/531, 436/535, 436/536, 435/7.94
• US Class Current: 435/7.94
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/54386 Analytical elements
• CPC Class Codes: G01N 33/54391 based on vertical flow

Abstract

The present invention provides a test carrier for the analytical determination of a component of a sample liquid with the help of immunological reagents having a conjugate layer (2, 12), which contains a soluble conjugate of a first immunological binding partner with a labelling, a porous fixing layer (3, 13), which contains a second binding partner in carrier-fixed form binding specifically with the first binding partner, and an optical detection layer (8, 18), wherein the fixing layer (3, 13) is constructed in such a manner that the suction speed with which the liquid is transported in it is so high that the time needed for the spreading out of the liquid in the fixing layer is considerably shorter than the incubation time which is needed for the completion of the specific binding reaction between the first binding partner and the second binding partner and the optical detection layer (8, 18) is so fixed on the test carrier (1, 11) that, in a first position, it is not in contact with the fixing layer (3, 13) making possible a liquid exchange but can be brought into a second position in which a liquid exchange between the fixing layer (3, 13) and the optical detection layer (8, 18) can take place.

The present invention also provides a process for the immunological analytical determination of a component of a sample liquid, as well as a process for the production of a fixing layer for a test carrier.

Claims

1. Test carrier useful in determination of an analyte in a liquid sample, comprising:

(i) a liquid absorbing first layer which contains a soluble conjugate of a first immunological binding partner and a label, the immunological binding partner of which specifically binds to said analyte to form a complex of analyte and conjugate;

(ii) a porous second layer in permanent fluid communication with said first layer and containing a solid phase bound analyte or analyte analogue which binds to said first immunological binding partner but does not specifically bind with said analyte, wherein said second layer absorbs liquid at a rate which spreads liquid therethrough in an amount of time less than an amount of time necessary for completion of a binding reaction between said solid phase bound analyte or analyte analogue and any of first immunological binding partner not bound to analyte, and

(iii) a third, liquid absorbing layer which contains a reagent which reacts with any conjugate of first immunological binding partner and label bound to said analyte to form an optically detectable signal, wherein said third layer is positioned on a flap means in said test carrier so that it is not in permanent contact with said second layer but may be brought into contact therewith as desired to permit liquid flow from said second layer to said third layer said contact permitting passage of liquid containing said complex of analyte and conjugate from said second layer into said third layer and penetration therein, to form said optically detectable signal.

2. Test carrier of claim 1, wherein said third layer is positioned in said carrier to permit contact from top or bottom of said second layer.

3. Test carrier of claim 1, wherein said second layer is characterized by a rate of liquid absorption greater than a rate of liquid absorption of said first layer.

4. Test carrier of claim 1, wherein a surface of said third layer faces said second layer, said surface being covered by a fourth layer which forms an optical barrier but is permeable to a liquid.

5. Test carrier of claim 1, wherein said second layer comprises a fiber composite structure.

6. Test carrier of claim 5, wherein said fiber composite structure is a fleece.

7. Test carrier of claim 5, wherein said fiber composite structure comprises synthetic resin fibers.

8. Test carrier of claim 5, wherein said fiber composite structure comprises glass fibers.

9. Test carrier of claim 8, wherein said fiber composite structure contains polyvinyl alcohol.

10. Test carrier of claim 1 or 5, wherein said second binding partner is bound to particles embedded in said second layer.

11. Test carrier of claim 1, wherein said particles consist of a synthetic resin.

12. Test carrier of claim 10, wherein said particles consist of inorganic material.

13. Test carrier of claim 10, wherein said particles have an average diameter of from about 0.01 u to about 5 u.

14. Test carrier of claim 13, wherein said particles have an average diameter of from about 0.3 u to about 1 u.

15. Test carrier of claim 1, further comprising a support layer to which said second layer is fixed, said second layer being defined by a sample application region at one end and an optical determination region at the other end, wherein said first layer is positioned in said sample application region and said third layer is positioned to permit contact with said second layer in said optical detection region.

16. Method for determination of an analyte in a liquid sample comprising contacting a liquid sample to a test carrier as in claim 1, incubating said sample to favor binding between (i) said analyte or a reaction product of said analyte and said conjugate and (ii) said conjugate and said second binding partner, and determining an optical change in said third layer as a measure of said analyte in said sample.

17. Process according to claim 16, wherein the liquid runs in the fixing layer parallel to its surface.